PA1-12528

antibody from Invitrogen Antibodies

Targeting: APP AD1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-12528

Provider

Invitrogen Antibodies

Product name

Anti-Amyloid Precursor Protein Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

A suggested positive control for this product is murine brain lysate.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Amyloid Precursor Protein Polyclonal Antibody(Product # PA1-12528) and a 120, 90kDa band corresponding to Amyloid Precursor Protein was observed in all cell lines and tissues tested except K-562, HEL 92.1.7, Mouse and Rat Skeletal Muscle which are reported to be negative. Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), K-562 (Lane 2), HEL 92.1.7 (Lane 3) and tissue extracts (30 ug lysate) of Mouse Brain (Lane 4), Mouse Skeletal Muscle (Lane 5), Rat Brain (Lane 6) and Rat Skeletal Muscle (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Amyloid Precursor Protein was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Amyloid Precursor Protein Polyclonal Antibody (Product # PA1-12528) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:3000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasm, Plasma membrane and Nuclear localization. Panel e represents merged image for K-562 cells showing no staining for APP. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.